LANES WORK

1. Collecting of Cells

± 10 mL Isotonic ± 10 mL ± 10 mL Mineral
Solution Aquadest Water

- Used to rinse for about 1 minute

- Entered into 3 different beaker glass

Sample Sample Sample

Solution Solution Solution

2. Lisis Cells and Protein Digestion

1,5 mL Sample Solution

- Taken with volumetric pipette

- Entered into micro centrifuge tube
- Centrifuge with 10.000 rpm in
microcentrifuge for 1 minute
- Separated by decantation process

Residue Filtrate

- Added 1,5 mL sample solution

- Centrifuge at 10.000 rpm until 1
minute
- Separated
- Repeated until 2 times

Residue Filtrate

- Added 1 mL buffer tris-EDTA

- Vortex microcentrifuge tube until 1
minute
- Observed the result

Lysis Cell solution

3. Precipitation of DNA

Lysis cell solution in

microcentrifuge tube

- Added 100 µL NaCl 2,5 M

- Mixtured
- Mised in another test tube carefully
- Added 1 mL cool ethanol
- Let in until 5 minutes

DNA dots between buffer and ethanol

boundaries (drifts likes white thread)
 OBSERVATION RESULT

Experiment Result
No. Procedure Prediction/Reaction Conclusion
Before After
1 Collecting of Cells  Isotonic  Isotonic sample solution: Turbidiry of solution Turbidiry based on
solution: turbid solution contain DNA experiment is
turbid  Aquadest sample solution: Isotonic solution >
solution turbid solution mineral > aquadest
 Aquadest:  Mineral water sample
colorless solution: turbid solution
solution
 Mineal water:
colorles
solution
2 Lisis Cells and Protein Digestion  Aquadest  Isotonic sample solution +
sample centrifuge = there is white
solution: precipitate and colorless
turbid  Separated:
 Isotonic Residue: white precipitate
sample Filtrate: colorless
solution:  +sample: turbid
turbid
 Mineral water  +centrifuge: white
sample precipitate and colorless
solution:  Residue + sample: turbid
turbid  +centrifuge: white
precipitate and colorless
 Separated:
Residue: white precipitate
Filtare: colorless solution
 +buffer tris EDTA: turbid
 +vortex: turbid

 Aqudest sample solution +

centrifuge: there is white
precipitate
 Separated:
Residue: white precipitate
Filtare: colorless solution
 Residue + sample: turbid
 +centrifuge: white
precipitate and colorless
 Residue + sample: turbid
  +centrifuge: white
precipiatte and colorless
 +buffer tris EDTA: turbid
 +vortex: turbid

 Mineral water sample

solution + centrifuge: white
precipitate and colorless
 Separated:
Residue: white precipitate
Filtare: colorless solution
 Residue + sample: turbid
 +centrifuge: white
precipitate and colorless
 Residue + sample: turbid
 +centrifuge: white
precipiatte and colorless
 +buffer tris EDTA: turbid
 +vortex: turbid
3 Precipitation of DNA  Lysis cell: Isotonic Solution DNA in Isotonic > Based on the
turbid  Lysis + NaCl: colorless mineral water > experiment, we
solution isotonic cell aqudest cannot state a
 NaCl:  +ethanol: colorless conclution because
colorless  Let in for 5 minutes: DNA is DNA is not isolated.
solution not isolated
 Cool ethanol:
Mineral Water
colorless
 Lysis + NaCl: colorless
solution
mineral water
 +ethanol colorless
 Let in for 5 minutes: DNA is
not isolated

Aquadest
 Lysis + NaCl: colorless
mineral water
 +ethanol colorless
 Let in for 5 minutes: DNA is
not isolated